This protocol provides the scientific community with a carefully developed guide describing, in 12 distinct steps, how to take structural images from data pre-processing, via statistical analysis, to the final interpretation of the significance maps. Key adaptations compared with the standard VBM workflow involve establishing a voxel-wise hemispheric correspondence, capturing the direction and degree of asymmetry and preventing a blurring of information across hemispheres. The workflow incorporates the most recent methodological developments, including high-dimensional spatial normalization and partial volume estimations. Although the protocol is primarily designed to enable relatively inexperienced users to conduct a voxel-based asymmetry analysis on their own, it may also be useful to experienced users who wish to efficiently adapt their existing scripts or pipelines.